Three sterol regulatory element-binding proteins (SREBP-1a, -1c, and -2) stimulate transcription of genes involved in synthesis and receptor-mediated uptake of cholesterol and fatty acids. Here, we explore the individual roles of each SREBP by preparing lines of Chinese hamster ovary (CHO) cells that express graded amounts of nuclear forms of each SREBP (designated nSREBPs) under control of a muristerone-inducible nuclear receptor system. The parental hamster cell line (M19 cells) lacks its own nSREBPs, owing to a deletion in the gene encoding the Site-2 protease, which releases nSREBPs from cell membranes. By varying the concentration of muristerone, we obtained graded expression of individual nSREBPs in the range that restored lipid synthesis to near physiologic levels. The results show that nSREBP-2 produces a higher ratio of synthesis of cholesterol over fatty acids than does nSREBP-1a. This is due in part to a selective ability of low levels of nSREBP-2, but not nSREBP-1a, to activate the promoter for squalene synthase. nSREBP-1a and -2 both activate transcription of the genes encoding stearoyl-CoA desaturase-1 and -2, thereby markedly enhancing the production of monounsaturated fatty acids. nSREBP-1c was inactive in stimulating any transcription at the concentrations achieved in these studies. The current data support the emerging view that the nSREBPs act in complementary ways to modulate the lipid composition of cell membranes.Cholesterol and fatty acids, the building blocks of cell membranes, are synthesized by regulated pathways in animal cells. Both pathways are influenced by a single family of transcription factors designated sterol regulatory element binding proteins (SREBPs), 1 whose concerted actions optimize the lipid content of membranes (reviewed in Brown and Goldstein (1)). Appropriate to their role in modulating membrane composition, the SREBPs are bound intrinsically to cell membranes.They are synthesized as long precursors of ϳ1150 amino acids with three domains. The NH 2 -terminal domain of ϳ480 amino acids is a classic basic helix-loop-helix-leucine zipper transcription factor. This is followed by a membrane attachment domain of ϳ80 amino acids and a COOH-terminal domain of ϳ590 amino acids that performs a regulatory function. The SREBPs are attached to membranes of the nuclear envelope and endoplasmic reticulum in a hairpin fashion with their NH 2 -terminal and COOH-terminal domains projecting into the cytosol and the membrane attachment domain projecting into the lumen (2).In order to reach the nucleus, the NH 2

A delicate balance of signals regulates cell survival. One set of these signals is derived from integrin-mediated cell adhesion to the extracellular matrix (ECM). Loss of cell attachment to the ECM causes apoptosis, a process known as anoikis. In searching for proteins involved in cell adhesion-dependent regulation of anoikis, we identified Bit1, a mitochondrial protein that is released into the cytoplasm during apoptosis. Cytoplasmic Bit1 forms a complex with AES, a small Groucho/transducin-like enhancer of split (TLE) protein, and induces cell death with characteristics of caspase-independent apoptosis. Cell attachment to fibronectin counteracts the apoptotic effect of Bit1 and AES. Increasing Bit1 expression enhances anoikis, while suppressing the expression reduces it. Thus, we have elucidated an integrin-controlled pathway that is, at least in part, responsible for the cell survival effects of cell-ECM interactions.

We have purified from hamster liver a second cysteine protease that cleaves and activates sterol regulatory element binding proteins (SREBPs). cDNA cloning revealed that this enzyme is the hamster equivalent of Mch3, a human enzyme that is related to the interleukin 1p3 con-